(3.235.139.152) 您好!臺灣時間:2021/05/11 06:16
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:許闊顯
研究生(外文):Kuo-Hsien
論文名稱:A群鏈球菌 M12蛋白之NH2端與COOH端重組蛋白在M12蛋白多倍體的形成與調理吞噬干擾作用之研究
論文名稱(外文):Study of the Group A streptococcus M12 NH2-terminal and COOH-terminal recombinant proteins in the multimer formation and opsonophagocytosis interference assays
指導教授:陳凌雲陳凌雲引用關係
指導教授(外文):Ling- Yun Chen
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生化暨生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:60
相關次數:
  • 被引用被引用:0
  • 點閱點閱:142
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
A型鏈球菌是是一種人類的致病菌,在全球每年會有大約1800萬人受到A型鏈球菌的影響,且會有51萬人死於A型鏈球菌所造成的疾病。A型鏈球菌所造成的疾病包含猩紅熱、風濕熱、風濕性心臟病、急性腎絲球腎炎等,其中風濕性心臟病是造成孩童以及年輕人得到心臟病的主要原因。在A型鏈球菌所有的毒性因子中,M蛋白是最重要的毒性因子。M蛋白是A型鏈球菌上的一種膜蛋白,在A型鏈球菌細胞膜上形成α-helical coiled coil結構。M蛋白幫助A型鏈球菌侵入宿主並且逃避宿主的免疫系統攻擊。本實驗室利用成人血清對許多不同隻A型鏈球菌做screen,實驗結果中發現在約57 kDa位置附近有一共同辨識蛋白,並且純化後具有調理干擾白血球之吞噬作用,經由MALDI-TOF鑑定是M12蛋白。因此本實驗製備M12的NH2端和COOH端重組蛋白做進一步確認,利用快速液相層析法(FPLC)以及調理吞噬干擾作用實驗來探討M12蛋白,發現M12的NH2端重組蛋白會形成聚集物,而COOH端重組蛋白則會形成12倍體,另外經由調理吞噬干擾實驗發現M12的COOH端重組蛋白能夠降低白血球對A型鏈球菌的吞噬作用,但卻無法完全抑制,原因在於成人血清為多株抗體,加入M12的COOH端重組蛋白無法拮抗所有認識M12蛋白的抗體,因而無法達到完全抑制白血球對A型鏈球菌吞噬的效果。

Group A Streptococcus(GAS) is a Human pathogen which causes 18.1 million cases disease and 0.51 million people dies. GAS causes severe disease including scarlet fever, rheumatic fever, rheumatic heart disease and acute glomerulonephritis, the rheumatic heart disease is the major reason which causes heart disease in the children and young people. For all of GAS toxicity factors, M protein is the most important toxicity factor. M protein is a membrane protein which have α-helical coiled coil structure on the plasma membrane. It helps GAS to attack the host and have resistance to the immune system. We use adult serum to test and screen different types of GAS and find that there is a common recognized band at about 57 kDa site, and it has the interference ability of the phagocytosis to the polymorphonuclear Leucocytes(PML). For the identification which is defined by MALDI-TOF is the type 12 M protein. We prepare the NH2-terminal and COOH-terminal recombinant M12 protein and use FPLC, opsonophagocytosis interference assays to confirm and study the M12 protein which found that M12 NH2-terminal recombinant protein is aggregate and the COOH-terminal recombinant protein is a dodecamer. The M12 COOH-terminal recombinant protein can decrease PML ability of GAS, but not complete inhibit the ability.
For the reason is that the adult serum is the polyclonal antibody and just put M12 COOH-terminal recombinant protein can not complete bind the all antibodies which can recognize the M12 protein. For the above description which is the reason for the M12 COOH-terminal recombinant protein can not complete inhibit the PML phagocytosis to the GAS.


目錄
縮寫表--------------------------------------------------2
圖次----------------------------------------------------3
附錄說明------------------------------------------------4
中文摘要------------------------------------------------5
英文摘要------------------------------------------------6
前言----------------------------------------------------7
實驗材料與方法------------------------------------------15
結果----------------------------------------------------29
討論----------------------------------------------------40
參考文獻------------------------------------------------43
附錄----------------------------------------------------46


1.Sriskandan S, Faulkner L, Hopkins P. Streptococcus pyogenes: Insight into the function of the streptococcal superantigens. Int J Biochem Cell Biol 2007,39:12-19.
2.Currie BJ. Group A streptococcal infections of the skin: molecular advances but limited therapeutic progress. Curr Opin Infect Dis 2006,19:132-138.
3.Olive C, Batzloff MR, Toth I. Lipid core peptide technology and group A streptococcal vaccine delivery. Expert Rev Vaccines 2004,3:43-58.
4.Carapetis JR, Steer AC, Mulholland EK, Weber M. The global burden of group A streptococcal diseases. Lancet Infect Dis 2005,5:685-694.
5.Johnson DR, Kaplan EL, VanGheem A, Facklam RR, Beall B. Characterization of group A streptococci (Streptococcus pyogenes): correlation of M-protein and emm-gene type with T-protein agglutination pattern and serum opacity factor. J Med Microbiol 2006,55:157-164.
6.Lancefield RC DV. The properties of T antigen extracted from group A hemolytic streptococci. J. Exp. Med. 1946,84:449-471.
7.Mora M, Bensi G, Capo S, Falugi F, Zingaretti C, Manetti AG, et al. Group A Streptococcus produce pilus-like structures containing protective antigens and Lancefield T antigens. Proc Natl Acad Sci U S A 2005,102:15641-15646.
8.Lizano S, Luo F, Bessen DE. Role of streptococcal T antigens in superficial skin infection. J Bacteriol 2007,189:1426-1434.
9.Jeng A, Sakota V, Li Z, Datta V, Beall B, Nizet V. Molecular genetic analysis of a group A Streptococcus operon encoding serum opacity factor and a novel fibronectin-binding protein, SfbX. J Bacteriol 2003,185:1208-1217.
10.Neal S, Beall B, Ekelund K, Henriques-Normark B, Jasir A, Johnson D, et al. International quality assurance study for characterization of Streptococcus pyogenes. J Clin Microbiol 2007,45:1175-1179.
11.Guilherme L, Kalil J. Rheumatic fever: the T cell response leading to autoimmune aggression in the heart. Autoimmun Rev 2002,1:261-266.
12.Stollerman GH. Rheumatic fever. Lancet 1997,349:935-942.
13.Brahmadathan KN, Gladstone P. Microbiological diagnosis of streptococcal pharyngitis: lacunae and their implications. Indian J Med Microbiol 2006,24:92-96.
14.Lancefield RC. The antigenic complex of Streptococcus haemolyticus. II. Chemical and immunological properties of the protein fractions. J. Exp. Med. 1928,47:469-480.
15.Phillips GN, Jr., Flicker PF, Cohen C, Manjula BN, Fischetti VA. Streptococcal M protein: alpha-helical coiled-coil structure and arrangement on the cell surface. Proc Natl Acad Sci U S A 1981,78:4689-4693.
16.Bisno AL, Brito MO, Collins CM. Molecular basis of group A streptococcal virulence. Lancet Infect Dis 2003,3:191-200.
17.Hosein B, McCarty M, Fischetti VA. Amino acid sequence and physicochemical similarities between streptococcal M protein and mammalian tropomyosin. Proc Natl Acad Sci U S A 1979,76:3765-3768.
18.Dale JB, Beachey EH. Epitopes of streptococcal M proteins shared with cardiac myosin. J Exp Med 1985,162:583-591.
19.Dale JB, Beachey EH. Multiple, heart-cross-reactive epitopes of streptococcal M proteins. J Exp Med 1985,161:113-122.
20.Giannakis E, Male DA, Ormsby RJ, Mold C, Jokiranta TS, Ranganathan S, Gordon DL. Multiple ligand binding sites on domain seven of human complement factor H. Int Immunopharmacol 2001,1:433-443.
21.Berggard K, Johnsson E, Morfeldt E, Persson J, Stalhammar-Carlemalm M, Lindahl G. Binding of human C4BP to the hypervariable region of M protein: a molecular mechanism of phagocytosis resistance in Streptococcus pyogenes. Mol Microbiol 2001,42:539-551.
22.Hollingshead SK, Fischetti VA, Scott JR. Complete nucleotide sequence of type 6 M protein of the group A Streptococcus. Repetitive structure and membrane anchor. J Biol Chem 1986,261:1677-1686.
23.Fischetti VA. Streptococcal M protein: molecular design and biological behavior. Clin Microbiol Rev 1989,2:285-314.
24.Dale JB, Beachey EH. Localization of protective epitopes of the amino terminus of type 5 streptococcal M protein. J Exp Med 1986,163:1191-1202.
25.Dale JB, Seyer JM, Beachey EH. Type-specific immunogenicity of a chemically synthesized peptide fragment of type 5 streptococcal M protein. J Exp Med 1983,158:1727-1732.
26.Jones KF, Fischetti VA. The importance of the location of antibody binding on the M6 protein for opsonization and phagocytosis of group A M6 streptococci. J Exp Med 1988,167:1114-1123.
27.Bronze MS, Beachey EH, Dale JB. Protective and heart-crossreactive epitopes located within the NH2 terminus of type 19 streptococcal M protein. J Exp Med 1988,167:1849-1859.
28.Beachey EH, Seyer JM. Protective and nonprotective epitopes of chemically synthesized peptides of the NH2-terminal region of type 6 streptococcal M protein. J Immunol 1986,136:2287-2292.
29.Jones KF, Khan SA, Erickson BW, Hollingshead SK, Scott JR, Fischetti VA. Immunochemical localization and amino acid sequences of crossreactive epitopes within the group A streptococcal M6 protein. J Exp Med 1986,164:1226-1238.
30.Strakova L, Motlova J, Jakubu V, Urbaskova P, Kriz P. Emergence of a novel macrolide-resistant Streptococcus pyogenes emm53 strain. Clin Microbiol Infect 2007,13:443-445.
31.Nakae M, Murai T, Kaneko Y, Mitsuhashi S. Drug resistance in Streptococcus pyogenes isolated in Japan. Antimicrob Agents Chemother 1977,12:427-428.
32.Schulze K, Medina E, Talay SR, Towers RJ, Chhatwal GS, Guzman CA. Characterization of the domain of fibronectin-binding protein I of Streptococcus pyogenes responsible for elicitation of a protective immune response. Infect Immun 2001,69:622-625.
33.Guzman CA, Talay SR, Molinari G, Medina E, Chhatwal GS. Protective immune response against Streptococcus pyogenes in mice after intranasal vaccination with the fibronectin-binding protein SfbI. J Infect Dis 1999,179:901-906.
34.Areschoug T, Carlsson F, Stalhammar-Carlemalm M, Lindahl G. Host-pathogen interactions in Streptococcus pyogenes infections, with special reference to puerperal fever and a comment on vaccine development. Vaccine 2004,22 Suppl 1:S9-S14.


QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
系統版面圖檔 系統版面圖檔